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Agriculturae Conspectus Scientii cus | Vol. 77 (2012) No. 1 (37-40)
ORIGINAL SCIENTIFIC PAPER
Summary
h e origins of biogenic amines are sound grapes, alcoholic fermentations, malol-actic fermentation and microbial activities during wine storage. h ese biologically produced amines are essential at low concentrations for optimal metabolic and phys-iological functions in animals, plants and micro-organisms. During alcoholic fer-mentation the degree of maceration is the i rst factor that af ects the extraction of compounds present in the grape skin, among them aminoacids, precursors of bio-genic amines. h e aim of the present work was to study the changes of the concentra-tion of biogenic amines in wines made from Vitis vinifera ‘Pinot noir’ from Plešivica (vintage 2009) produced with classical maceration, cold maceration and use of sur lie method. Biogenic amines were quantii ed using a reversed-phase high performance liquid chromatography (HPLC) with l uorescence detection at er pre-column deri-vatization with o-phthalaldehyde (OPA). In ‘Pinot noir’ wines tested, histamine was the most abundant biogenic amine followed by tryptamine and 2-Phenylethylamine. Total amount of biogenic amines ranged from 8.72 mg/L in wines made with classical maceration up to 9.34 mg/L in sur lie wines. In summary, from the results obtained in this study, it can be concluded that sur lie technology can inl uence the formation of biogenic acids since the release of amino acids is probably more pronounced in wines aged with lees and stirred weekly. No signii cant dif erences were found in the con-centration of biogenic amines in relation to the used maceration process.
Key words
biogenic amines, ‘Pinot noir’, maceration, sur lie
Concentration of Biogenic Amines in
‘Pinot Noir’ Wines Produced in Croatia
Ana JEROMEL
1 ( )Karin KOVAČEVIĆ GANIĆ
2Stanka HERJAVEC
1Marin MIHALJEVIĆ
1Ana Marija JAGATIĆ KORENIKA
1Ivana RENDULIĆ
3Marijana ČOLIĆ
41 University of Zagreb, Faculty of Agriculture, Svetošimunska 25, 10000 Zagreb
e-mail: [email protected]
2 University of Zagreb, Faculty of Food Technology and Biotechnology,
Pierottijeva 6, 10000 Zagreb, Croatia
3 Zagreb County, Ulica grada Vukovara 72, 10000 Zagreb, Croatia 4 student, University of Zagreb, Faculty of Agriculture,
Svetošimunska 25, 10000 Zagreb, Croatia Received: March 4, 2011 | Accepted: May 31, 2011 ACKNOWLEDGEMENTS
Agric. conspec. sci. Vol. 77 (2012) No. 1
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Ana JEROMEL, Karin KOVAČEVIĆ GANIĆ, Stanka HERJAVEC, Marin MIHALJEVIĆ, Ana Marija JAGATIĆ KORENIKA, Ivana RENDULIĆ, Marijana ČOLIĆIntroduction
Biogenic amines (BA) are nitrogenous low molecular weight organic bases that can have an aliphatic, aromatic or heterocyclic structure. h ey are widely present in food, especially in ferment-ed food, mostly as a consequence of the decarboxylation of their free precursor amino acids (Vincenzini et al., 2009). h e amines histamine, tyramine, putrescine, cadaverine, 2-phenethylamine, agmatine and tryptamine originate from the precursor amino acids histidine, tyrosine, ornithine, lysine, phenylalanine, argi-nine and tryptophane (Bauza et al., 1995). Musts from grape usu-ally contain low concentration of BA, almost entirely represented by spermidine and putrescine. h is diamine is reported to be synthesized by the vine in response to stress conditions (Halasz et al. 1994). Wines are usually characterized by a signii cantly higher content of BA, red wines compared to white being gener-ally characterized by higher BA content. Soul eros et al. (2007) reported up to 2.11 mg/L histamine, 3.65 mg/L tyramine and 5.23 mg/L putrescine in red wines from Greece, while Marcobal et al. (2005) found in red Spanish wines 3.62 mg/L histamine, 1.40 mg/L tyramine and 7.06 mg/L putrescine. Generally, the toxic dose in alcoholic beverages is considered to be between 8 and 20 mg/L for histamine, 25 to 40 mg/L for tyramine, but as little as 3 mg/L phenylethylamine can cause negative physiologi-cal ef ects (Soul eros et al., 1998). A wide variety of viticultural and oenological factors may have an impact on the levels of bio-genic amines produced in wine. While some factors can increase the precursor amino acid concentration in the grape and wine (grape variety, geographical region, vintage, grape skin macera-tion, ageing practices), other factors inl uence diversity, growth, decarboxylase activity that can potentially produce the biogenic amines. Some amines, such as putrescine and other polyamines, may be already present in grape berries (Bover-Cid et al., 2006). ‘Cabernet sauvignon’ was found to have high concentration of putrescine, cadaverine and spermidine in the pericarp of the ber-ries (Glória et al., 1998). Putrescine, cadaverine and spermidine have been also found in high concentration in the seeds of grape berries (Kiss et al., 2006). h erefore, putrescine concentration in wine may be inl uenced more by geographical region and grape variety than by winemaking practices. Biogenic amines are also dependent on grape variety and vine nutrition, which will de-termine the concentration and composition of precursor amino acids in grape must (Soul eros et al., 1998). Among the factors that have been suggested as favouring the abundance of amines in wine, some winemaking practices, such as nutrient addition and duration of wine contact with both grape skins and yeast lees, seem to play a major role because they can directly af ect the content of the precursor amino acids of BA (Vincenzini et al., 2009). Grape skin maceration promotes extraction of grape components such as phenolic compounds, proteins, amino acids and polysaccharides. Soleas et al. (1999) found no correlation between length of skin contact and concentration of biogenic amines, while Martín-Alvarez et al. (2006) found that the du-ration of skin macedu-ration signii cantly af ects biogenic amine content in wine.
During ageing of the wine with lees, amino acids are released by yeast and bacterial autolysis. According to Alcaide-Hidalgo et al. (2007) this release of amino acids is more pronounced in wines aged with lees and stirred weekly. h erefore, precaution
should be taken when wines are aged with lees, since this in-creases the risk of the formation of biogenic amines by residual decarboxylase activities of the lactic acid bacteria responsible for malolactic fermentation.
h e aim of the present work was to study the changes of the content of biogenic amines in wines made from grapevine vari-ety ‘Pinot noir’ (Vitis vinifera L.) from Plešivica in vintage 2009 and produced with classic maceration, cold maceration and use of sur lie method.
Materials and methods
Samples and fermentationGrape from ‘Pinot noir’ produced in northwest Croatia, Plešivica winegrowing district was harvested at optimum maturity in 2009. Grape washarvested manually, placed in plastic boxes and transported to the vinery. Grape was destemmed, crushed and then transferred into stainless steel tanks for maceration. h e cold-maceration was carried out controlling the skin contact time for i ve days at temperature below of 15°C by using dry ice. At er the cold-maceration period was completed, the tempera-ture of the tanks was let to rise up to 25°C to allow starting the alcoholic fermentation. Classic fermentation was carried out on controlling the skin contact for eight days at 25°C. At er this, the mash was drawn of to remove the skins and other solid parts, and the free-run musts were transfer to 225 L barrique barrels and let to i nish the fermentation under the same conditions. Alcoholic fermentation was carried out using of Uvaferm 229 starter culture (Lallemand). At the end of fermentation all wines were inoculated with commercial pure strain culture Uvaferm Alpha (Lallemand) for conduction of malolactic fermentation. At er that, the wine was racked, total sulphur dioxide adjusted to 50 mg/L and let to mature in cellar conditions. Sur lie wines were aged by weekly stirring of lees during six months period, when all the wines (from i ve repetition) were taken for analy-sis. Basic chemical analyses of must and wine were done using methods proposed by O.I.V. (2001).
HPLC determination of biogenic amines
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Concentration of Biogenic Amines in ‘Pinot Noir’ Wines Produced in Croatiaa Varian Pro Star Solvent Delivery System 230 (Varian, Walnut Creek, USA) and a Fluorescence detector Varian ProStar 363 (Varian, Walnut Creek, USA), using a reversed-phase column Pinnacle II C-18 column (Restek, USA)(150 x 3.9 mm, 5μm i.d.). Chromatographic conditions were: solvent (A): 0.05 M sodium acetate buf er adjusted to pH 6.6 / tetrahydrofuran (96:4); sol-vent (B) 100% methanol at a l ow rate of 1.2 mL/min. h e elu-tion was performed with a gradient starting at 100% B to reach 53% B at 2.5 min, 70% B at 7.5 min and 100% B at 15 min, and becoming isocratic for 10 min. Detection was carried out using 340 nm and 420 nm as excitation and emission wavelengths re-spectively. h e content of each analyte was obtained by direct interpolation of the peak area in the correspondent linear cali-bration curve. Because certain biogenic amines are in salt form, the weight of the salt was taken into account when determin-ing the true weight of the biogenic amine. h e data acquisition and treatment were conducted using the Star Chromatography Workstation Version 5 sot ware.
Statistical analysis
One-way analysis of variance (ANOVA) and Least Signii cant Dif erence (LSD) comparison test of SAS (SAS Institute, Cary, NC, USA) were used to interpret statistical dif erences in means, if any, at the P < 95% coni dence level.
Results and discussion
As it can be seen from the data shown in Table 1 there was no pronounced dif erence in basic chemical composition of tested wines. Signii cant dif erence was noted in ash, total phenols and in l avan-3-ols concentration mainly due to used fermen-tation technique. h e importance of wine lees in this context comes from the fact that lees can adsorb phenolic compounds (Mazauric and Salmon, 2005) and release to wine both phenolic compounds (Somers et al., 1987) and enzymes (at er autolysis) that can modify the phenolic fraction concentration (Ibern-Gómez et al., 2000).
h e concentration of biogenic amines produced in wine largely depends on the abundance of amino acid precursors in the medium since biogenic amines increase with an increase in amino acids. Amino acid concentration may be inl uenced by vinii cation methods, grapevine variety, geographical condi-tions (ecological condicondi-tions) and vintage (Soul eros et al., 1998). According to our results vinii cation method signii cantly in-l uenced the concentration of some biogenic amines (Tabin-le 2). Wines aged on the lees had signii cantly higher concentration of histamine, 2-phenylethylamine and tryptamine while there was no dif erence between wines produced with classical maceration and cold maceration. Martin-Álvarez et al. (2006) let their wines in contact with lees for two months at er alcoholic fermenta-tion and noticed increase in methylamine and putrescine while tyramine concentration was lower. Our results are not in agree-ment with that stateagree-ment, probably because wines were analyzed at er longer ageing period. Various research groups (Gerbaux and Monamy 2000, Landete et al., 2005) noticed that at er the initial increase in biogenic amines during storage, a general de-crease or stabilization in concentration could be observed. But Coton et al. (1998) found out that even when no more cultur-able cells were detectcultur-able, hystidine decarboxylase could still be
active-thus biogenic amines can be produced during aging. In ‘Pinot noir’ wines tested, histamine was the most abundant bio-genic amine followed by tryptamine and 2-Phenylethylamine. In general, putrescine is the major biogenic amine found in wines and putrescine producing capability may be considered wide-spread among lactic acid bacteria strains of oenological interest (Moreno-Arribas et al., 2003). According to Alcaide-Hidalgo et al. (2007) putrescine content of the wines aged with lees increas-es during ageing, although it remains stable in those aged with-out lees. As shown in Table 2 putrescine concentration in ‘Pinot noir’ was relatively similar and low probably because malolactic fermentation was conduced by commercial pure strain culture Uvaferm Alpha. In summary, from the results obtained in this study, it can be concluded that sur lie technology can inl uence the formation of biogenic acids since the release of amino acids is probably more pronounced in wines aged with lees and stirred weekly. No signii cant dif erences were found in the concen-tration of biogenic amines in relation to the used maceration process. Our results showed no toxic dose in ‘Pinot noir’ wines tested ranging between 3.26-3.49 mg/L for histamine, 0.17-0.22 mg/L for tyramine, and 1.35 -1.69 mg/L for phenylethylamine. h e toxic dose of these biogenic amines in alcoholic beverages is
Compounds Classic maceration
Cold maceration
Sur lie LSD5%
Alcohol (vol%) 13.68 13.71 13.62 n.s. Residual sugar (g/L) 2.5 2.9 2.2 n.s. Total extract (g/L) 26.54 26.38 27.94 n.s.
Total acidity (g/L)* 6.9 6.8 6.5 n.s
Volatile acidity (g/L)** 0.62 0.60 0.69 n.s.
pH 3.62 3.69 3.71 n.s.
Ash (g/L) 2.36a 2.28a 2.49b 0,08
Total phenols (mg/L, galic acid equivalents)
2047.11a 2098.23b 2157.51c 19.95
Flavan-3-ols (mg/L, catechin equivalents)
145.56a 147.23a 195.11b 8.45
Different letters beside the mean of a compound denote a significant difference among treatments; n.s.: not significant; *as tartaric acid; **as acetic acid
Biogenic amine Classic maceration
Cold
maceration Sur lie LSD 5% Histamine 3.26a±0.06 3.36a±0.09 3.49b±0.06 0.12 Tyramine 0.17±0.01 0.20±0.01 0.22±0.02 n.s.
Putrescine 0.39±0.09 0.40±0.08 0.46±0.07 n.s
Cadaverine 0.32±0.05 0.34±0.06 0.40±0.05 n.s
2-Phenylethylamine 1.35a±0.13 1.47a±0.15 1.69b±0.19 0.11
Spermidine 0.52±0.11 0.59±0.09 0.61±0.09 n.s
Tryptamine 2.55a±0.10 2.74b±0.12 2.89b±0.17 0.09 Spermine 0.06±0.01 0.07±0.01 0.08±0.02 n.s
Serotonine 0.08±0.05 0.11±0.04 0.10±0.04 n.s
Σ 8.72a 8.86a 9.45b 0.19
Table 1. Determined chemical characteristics of wines ‘Pinot
noir’ produced in 2009 according to dif erent treatment
Table 2. Concentration of determined biogenic amines in
Agric. conspec. sci. Vol. 77 (2012) No. 1
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Ana JEROMEL, Karin KOVAČEVIĆ GANIĆ, Stanka HERJAVEC, Marin MIHALJEVIĆ, Ana Marija JAGATIĆ KORENIKA, Ivana RENDULIĆ, Marijana ČOLIĆconsidered to be between 8 and 20 mg/L for histamine, 25 and 40 mg/L for tyramine, but as little as 3 mg/L phenylethylamine can cause negative physiological ef ects (Soul eros et al., 1998).
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